Relating interindividual differences in cerebral organoid models to longitudinal infant brain growth and cognitive development

项目总结/摘要 诱导多能干细胞（iPSC）和人类大脑类器官准备提供洞察如何 神经发育障碍患者的大脑发育变化。虽然分子保真度 皮质类器官已相对于主要胎脑参考数据集进行了评估，但尚不清楚如何进行评估。 皮质类器官表型预测了出生后纵向脑生长的个体间差异。 确定哪些体外表型与体内性状相关将有助于解释实验 实施类器官以了解神经发育障碍的机制。为了检验这种关系， 是从充分表征的个体衍生iPSC系群体所必需的。该提案使用iPSC- 来自婴儿脑成像研究（IBIS）参与者的脑类器官，以测试 类器官神经上皮芽结构和基因表达表型与体内脑发育之间的关系 和认知行为测量。具体地说，该提案使用了从5个细胞中产生的17个iPSC细胞系。 没有神经精神疾病家族史的参与者，9名有ASD兄弟姐妹的参与者， 和3名ASD参与者，每个参与者在6、12和/或24个月大时进行至少2次脑部MRI扫描。所有 参与者有Mullens早期学习量表，Vineland适应行为和自闭症诊断 24月龄时的观察量表评分。产生每个iPSC系的3个克隆。为了识别- iPSC供体之间和同一个体的iPSC克隆内的个体差异，我们将评估 单细胞基因表达和类器官结构的分化。单细胞RNA测序将 在神经上皮芽形成（分化后14天）和神经元成熟（84天）后进行 分化后）。完整的类器官免疫标记将在这些时间点和更早的时间点完成。 神经发生（分化后56天）。完成后，该项目将确定优势， 大脑类器官作为人类皮质发育的可重复预测模型的局限性。这 该提案旨在利用严格的课程，辅导，网络和技术培训， 位于查佩尔山的北卡罗来纳州大学，并通过IBIS合作者。通过完成这项建议， 申请人将有适当的技术培训和沟通技巧，成为独立的 在神经发育领域拥有强大专业网络的研究者。

PROJECT SUMMARY/ABSTRACT Induced pluripotent stem cells (iPSCs) and human cerebral organoids are poised to provide insight into how brain development changes in individuals with neurodevelopmental disorders. Though the molecular fidelity of cortical organoids have been assessed relative to the primary fetal brain reference datasets, it is unknown how well cortical organoid phenotypes predict inter-individual differences in postnatal longitudinal brain growth. Identifying what in vitro phenotypes are related to in vivo traits will be useful for interpreting experiments implementing organoids to understand mechanisms of neurodevelopment disorders. To test this relationship, it is necessary to derive a population of iPSC lines from well-characterized individuals. This proposal uses iPSC- derived cerebral organoids from participants in the Infant Brain Imaging Study (IBIS) to test the relationship between organoid neuroepithelial bud structure and gene expression phenotypes to in vivo brain development and cognitive behavioral measurements. Specifically, this proposal uses 17 iPSC lines generated from 5 participants without a family history of neuropsychiatric disorders, 9 participants who had a sibling with ASD, and 3 participants with ASD, each with at least 2 brain MRI scans at 6, 12 and/or 24 months of age. All participants have Mullens Scale of Early Learning, Vineland Adaptive Behavior and Autism Diagnostic Observation Scale score at 24 months of age. 3 clones of each iPSC line were generated. To identify inter- individual differences between iPSC donors and within iPSC clones from the same individual, we will assess single cell gene expression and organoid structure across the differentiation. Single cell RNA sequencing will be performed after neuroepithelial buds form (14 days post differentiation) and neuronal maturation (84 days post differentiation). Intact organoid immunolabeling will be completed at these time points and earlier in neurogenesis (56 days post differentiation). When completed, this project will identify the strengths and limitations of cerebral organoids as reproducible, predictive models of human cortical development. This proposal seeks to leverage the rigorous coursework, mentorships, networks and technical training available at the University of North Carolina at Chapel Hill and through IBIS collaborators. By completing this proposal, the applicant will have the appropriate technical training and communication skills to become an independent investigator with a strong professional network in the field of neurodevelopment.